松脂醇二葡萄糖苷大鼠胆汁内排泄研究

Study on bile excretion of pinoresinol diglucoside of rats

关键词:  松脂醇二葡萄糖苷  液质联用  胆汁排泄  代谢产物

Key words:  pinoresinol diglucoside  LC-MS/MS  biliary excretion  metabolite

摘要:

[目的] 建立液质联用方法，同时测定胆汁中松脂醇二葡萄糖苷和其代谢产物松脂醇单葡萄糖苷的含量，研究两者在胆汁内排泄特征。[方法] 大鼠灌胃给药松脂醇二葡萄糖苷的生理盐水溶液，并按照设定的时间点收集胆汁。以华法林单体为内标，建立液质联用分析方法，测定灌胃给药后大鼠胆汁中松脂醇二葡萄糖苷和其代谢产物松脂醇单葡萄糖苷的含量变化情况。[结果] 松脂醇二葡萄糖苷和松脂醇单葡萄糖苷在8~5 000 ng/mL范围内线性关系良好，日内精密度和日间精密度及其他方法学考察项RSD均符合生物样本定量要求。给药后松脂醇二葡萄糖苷和松脂醇单葡萄糖苷的累积排泄量在给药之后不断增长，到6 h后保持一个平稳状态；排泄速率在4 h时到达顶峰，之后迅速降低，6 h后保持一个较低的状态，实验后期大鼠排泄率有上升趋势。[结论] 该方法符合生物样品检测要求，可用于松脂醇二葡萄糖苷和松脂醇单葡萄糖苷的同时定量测量。

Abstract:

[Objective] To establish a method for simultaneous determination of pinoresinol diglucoside and its metabolites pinoresinol singleglucoside in rat bile by liquid chromatography-mass spectrometry (LC-MS/MS) and to study the excretion of two compound from rat bile. [Methods] Bile samples were collected according to the designed time periods after intragastric administration. The content of pinoresinol diglucoside, with the warfarin prime as internal standard, and it's metabolites in rat bile were then analyzed using the established LC-MS/MS method. [Results] Calibration curves were generated over a concentration range of 8~5 000 ng/mL for pinoresinol diglucoside and pinoresinol singleglucoside, respectively. Interday, intra-day and other methodology study RSDs were in accordance with the requirements of the biological samples quantitatively. Biliary accumulated excretion after administration were growing continually, to maintain a steady state after 6 hours; discharge rate in 4 hours to reach the top, afterdischarge rate falling rapidly, after 6 hours in a low condition,rats excretion rate has a rasing trend in the late experiment.[Conclusion] This method meets the requirements for the quantitative determination of biological samples and can be used forquantitative analysis of pinoresinol diglucoside and it's metabolites pinoresinol singleglucosidein rat bile.